Objective:To study the effect of GnRH analog triptorelin in resensitizing cisplatin-resistant human ovarian canc- er cells and to discuss the related mechanism.Methods:Cisplatin-resistant human ovarian cancer cell line OVCAR-3/CDDP was established in vitro.MTT assay was used to assess the inhibitory effects of triptorelin,cisplatin alone or a combination of both on OVCAR-3/CDDP cells.Flow cytometry was employed to observe the expression changes in epidermal growth factor receptor (EGFR)in different groups.Results:The drug restant index of OVCAR-3/CDDP cells was 13.42.The resensitizing fold of cisplatin combined with triptorelin was 3.80.The expression of EGFR had the most prominent decrease in OVCAR-3/CDDP cells in the combination group.Conclusion:Triptorelin can partially resensitize cisplatin-resistant OVCAR-3/CDDP cells,which might be related to the down-regulation of EGFR.

Objective To established cardiac-specific transgenic mice of the cTnC D145E and cTnCG159D and compare the HCM and the DCM.Methods The cTnCD145E and cTnCG159D were generated by site-directed mutagenesis and the transgenic plasmids were constructed by insertion of the mutant genes under the control of α-MHC, which is a myocardium specific promoter.The transgenic mice were generated by microinjection and were all maintained on a C57BL/6J genetic backgroud .The cardiac structure and function of the transgenic mice were compared and analysized by echocardiographic and pathological observation at different ages .Results The cTnCD145E and cTnCG159D transgenic mice were established and developed to HCM and DCM, respectively, with aging.The left ventricular end-systolic volume (ESV) and left ventricular end-diastolic volume ( EDV) decreased and ejection fraction ( EF) and left ventricular end-systolic posterior wall thickness (ESPWT) increased in the cTnCD145E transgenic mice, while EDV and ESV increased and EF and ESPWT decreased in the cTnCG159D transgenic mice at 12 months of age.Conclusions Cardiac-specific human cTnCD145E transgenic mice showed HCM phenotypes , and cardiac-specific human cTnC G159D transgenic mice showed DCM phenotypes , which can be used as different models for comparative study of the pathogenesis of cardiomyopathy .

To investigate whether epigenetic alterations in the insulin-like growth factor-II (IGF-II) gene that cause differential transcription or expression are correlated with onset and severity of hepatocellular carcinoma (HCC). Patient-matched specimens of HCC, paracancerous, and non-cancerous tissues were collected from 40 primary liver cancer patients. Epigenetic alterations in the promoter (P3) sequence of the IGF-II gene were analyzed by methylation-specific PCR (MSP) and IGF-II transcription was measured by RT-PCR. IGF-II protein expression and clinicopathological features were assessed by immunohistochemistry and microscopic observation. The rate of IGF-II P3 methylation was significantly lower in HCC tissues (0%) than in paracancerous tissues (vs. 47.5%; x2 = 24.918, P less than 0.001) and non-cancerous tissues (vs. 100%; x2 = 80.000, P less than 0.001). IGF-II mRNA expression was significantly higher in HCC tissues (100%) than in paracancerous tissues (vs. 52.5%; x2 = 24.918, P less than 0.001) and non-cancerous tissues (vs. 0%; x2 = 80.000, P less than 0.001). IGF-II protein expression was significantly higher in HCC tissues (82.5%) than in paracancerous tissues (vs. 45.0%; x2 = 12.170, P less than 0.001) and non-cancerous tissues (vs. 0%; x2 = 56.170, P less than 0.001). IGF-II overexpression in HCC was significantly associated with degree of differentiation, extent of infiltrated serosa, size of tumor, and HBV-positive infection status. Epigenetic alterations in the IGF-II gene regulate its transcription and expression and are closely associated with HCC development and progression.
Adult ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; CpG Islands ; genetics ; DNA Methylation ; Epigenesis, Genetic ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Insulin-Like Growth Factor II ; genetics ; metabolism ; Liver ; metabolism ; pathology ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Middle Aged ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; metabolism ; Transcription, Genetic

OBJECTIVETo investigate the effect of miRNA silencing HIF-1α gene on the proliferation of HepG2 cells.

METHODSThe eukaryotic expression plasmids of HIF-1α miRNA and report gene containing hypoxia-reponse element were constructed and transfected into HepG2 cells. The expressions of HIF-1α gene and protein were determined by real time-PCR and Western blotting. The expressions of HIF-1α, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) were quantitatively detected by ELISA. The alterations of cell cycles and apoptosis rate were quantitatively measured by flow cytometry and Annexin V-FITC/PI double dyeing assay.

RESULTS72 h after transfection the down regulations of HIF-1α mRNA and protein were 87% and 56% respectively, and the decrease of target gene was 46% in the report gene, 54% in VEGF and 36% in Ang-2, respectively. The apoptotic ratio of HepG2 cells was 22.46+/-0.61% (P < 0.01). The cell cycle changed greatly at the ratio of G1 (61.49+/-1.12%) and S (22.40+/-0.58%, P < 0.01). After being combined with doxorubicin, the apoptotic ratio increased to 36.99+/-0.88% and the ratios of G1 and S phases were upregulated to 65.68+/-0.91% and 19.47+/-1.34% respectively.

CONCLUSIONSHIF-1α miRNA or / and doxorubicin can regulate the growth cycles of HepG2 cells, promote the cell apoptosis and inhibit the cell proliferation.

Apoptosis ; Cell Cycle ; Cell Proliferation ; Gene Silencing ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; MicroRNAs ; genetics ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo analysis the complications of coronary rotational atherectomy and evaluate the safety of this procedure.

METHODA total of 250 rotational atherectomy cases from April 1994 to February 2012 were screened retrospectively and 22 cases patients (8.8%) with rotational atherectomy-related complications were included in this analysis.

RESULTSAmong these 22 patients, all lesions were either type B2 or C calcified lesions as evidenced by coronary angiography. After the rotation procedure, there were seven cases (2.8%) with slow reflow and two (0.8%) cases with no reflow. Seven cases (2.8%) developed severe coronary spasm and two cases (0.8%) had sinus bradycardia. Coronary dissection occurred in two cases (0.8%), while one case (0.4%) had coronary perforation and cardiac tamponade. Burr entrapment happened in one case (0.4%). There was no malignant arrhythmia, acute myocardial infarction, emergent coronary artery bypass graft or device related death during and post procedure. Comparison with baseline data, the concentration of CK-MB elevated significantly after the rotational atherectomy [(31.2 ± 4.8) mmol/L vs. (11.4 ± 6.5) mmol/L, P < 0.05].

CONCLUSIONCoronary rotational atherectomy is safe and procedure-related complications are rare.

Aged ; Aged, 80 and over ; Atherectomy, Coronary ; adverse effects ; methods ; Female ; Humans ; Intraoperative Complications ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo compare the efficacy and safety between the interventional and conservative treatment options for borderline vulnerable plaque lesion in acute coronary syndrome (ACS) patients by intravascular ultrasound (IVUS).

METHODSA total of 100 ACS patients [78 male, age 43 - 74 (60.4 ± 14.1) years] undergoing coronary angiography (CAG) with borderline lesion (coronary artery stenosis between 50% - 70%) were enrolled in May 2007 to February 2009, who were randomly divided into PCI group (50 patients) and conservative therapy group (50 patients). According to minimal lumen area (MLA) detected by IVUS, patients were further divided into MLA ≥ 4.0 mm(2) sub-group and MLA < 4.0 mm(2) sub-groups. Outcomes during hospitalization and after 10 - 12 month follow-up were compared.

RESULTSIVUS was performed in 40 patients at 10 - 12 months post PCI, there was no in-stent thrombosis and the extent of stent neointimal hyperplasia was comparable as at the time of immediately post PCI. IVUS was performed in 35 patients at 10 - 12 months post conservative therapy, IVUS results showed that MLA increased significantly [(7.32 ± 1.42) mm(2) vs. (4.98 ± 0.89) mm(2), P < 0.01], while plaque area [(7.70 ± 2.09) mm(2) vs. (10.01 ± 2.55) mm(2), P < 0.05], plaque burden [(55.94 ± 8.36)% vs. (67.97 ± 9.36)%] and low echo area [(4.08 ± 0.80) mm(2) vs. (2.27 ± 0.79) mm(2)] were significantly decreased at follow up compared to those as baseline (all P < 0.01). There was one patient in PCI group with MLA ≥ 4.0 mm(2) developed acute in-stent thrombosis in left anterior descending artery two days after the procedure and 9 patients in conservative therapy and MLA < 4.0 mm(2) group received PCI due to recurrent angina pectoris during follow-up.

CONCLUSIONSFor the borderline lesion with MLA ≥ 4.0 mm(2) detected by IVUS, adequate medication could effectively attenuate and or reverse the plaque progression and stabilize plaque.

Acute Coronary Syndrome ; drug therapy ; therapy ; Adult ; Aged ; Catheter Ablation ; Coronary Angiography ; Female ; Humans ; Male ; Middle Aged ; Plaque, Atherosclerotic ; diagnosis ; Treatment Outcome ; Ultrasonography, Interventional

Objective:To investigate the characteristics of pregnancy with pre-excitation syndrome and its influence on pregnancy outcomes.Methods:A retrospective analysis was made on the clinical data of 62 cases of pregnancy complicated with pre-excitation syndrome in Beijing Anzhen Hospital from Jan. 2008 to Dec. 2008. According to whether there was a supraventricular tachycardia (SVT) in pregnancy, they were divided into two groups. There were 16 pregnant women in the SVT seizure group during pregnancy, and 46 pregnant women in no SVT seizure group, with a multi-disciplinary comprehensive diagnosis and treatment model. SPSS software was used to analyze the data and compare the gestational age, age and weight of the newborn, and then compare the pregnancy outcomes.Results:(1) The total number of deliveries in Beijing Anzhen Hospital during the study period was 21 786, and the patients with pregnancy combined with pre-excitation syndrome account for 0.28% (62/21 786). (2) Totally 44 patients (71%, 44/62) were diagnosed with pre-excitation syndrome before pregnancy, and 18 patients (29%, 18/62) were diagnosed for the first-time during pregnancy. Among patients diagnosed with pre-excitation syndrome before pregnancy, 16 patients (36%, 16/44) had seizures before pregnancy, 28 patients (64%, 28/44) had asymptomatic before pregnancy, and 4 of asymptomatic patients had SVT during pregnancy. (3) Of the 16 pregnant women in the SVT seizure group during pregnancy, 2 patients (2/16) had SVT episodes in the first trimester, 5 patients (5/16) had SVT episodes in the second trimester, 9 patients (9/16) had SVT episodes in the third trimester. In the SVT seizure group, 8 patients (8/16) had SVT episodes before pregnancy, and 8 patients (8/16) had no SVT episodes before pregnancy. There were 46 patients in the SVT seizure-free group during pregnancy, including 9 patients with SVT attacks before pregnancy and 37 patients without SVT before pregnancy. (4) Compared with the pregnant women in the SVT seizure group, the age, weight gained during pregnancy, delivery gestation week, newborn weight, and the time of the first and second labors were not statistically different between the two groups of pregnant women (all P>0.05). However, the total duration of labor in the SVT seizure group during pregnancy was shorter and pre-pregnancy weight was lower (all P<0.05). The rate of cesarean section in pregnant women with SVT attack was 12/16, and the rate of cesarean section in pregnant women without SVT was 50% (23/46; P=0.051). No pregnant woman had an arrhythmia during delivery. Conclusions:SVT episode in patients during pregnancy most occurs in the third trimester. Patients who are asymptomatic before pregnancy may also have SVT during pregnancy. Pre-excitation syndrome patients with SVT attacks during pregnancy increase adverse pregnancy outcomes. Multidisciplinary comprehensive management could effectively control pregnant women with pre-excitation syndrome, effectively reduce the occurrence of serious arrhythmia risk events during pregnancy, so that most patients could get good pregnancy outcomes.

Objective To study the effects of fluorocitrate (FC),astrocytic metabolic inhibitor,on early brain injury after subarachnoid hemorrhage (SAH)in rats so as to explore the mechanism of early brain injury mediated by astrocyte after SAH.Methods Thirty six SD male rats were randomly divided into sham group,SAH 3 d group and SAH 3 d+FC group,with 12 rats in each group.The model of SAH was established by an endovascular perforation technique. FC was injected into the lateral ventricle. Three days after SAH, the expressions of GFAP,Iba-1 and TNF-α were detected using immunohistochemistry in all rats.The expression and phosphorylation of NF-κB in the nucleus were detected by Western blot.The expressions of TNF-α,IL-1βand IL-6 were detected by ELISA.Cell apoptosis was detected by TUNEL.Results Neuronal swelling,nuclear anomalies, disorganization of micrangium,abnormality of endothelial cells appeared in SAH 3 d group,but not in sham group. The expressions of NSE,GFAP and Iba-1 were significantly increased in SAH 3 d group compared with those in sham group (all P＜0.05).The number of apoptotic cells was increased.The expression and phosphorylation of NF-κB were significantly increased.The expressions of TNF-α,IL-1β and IL-6 were significantly increased.However, neuronal injuries were alleviated by injecting FC.The expressions of NSE,GFAP and Iba-1 in SAH 3 d+FC group were inhibited.The number of apoptotic cells in SAH 3 d+FC group was decreased compared with that in SAH 3 d group.The expression and phosphorylation of NF-κB were decreased by FC.The expressions of TNF-α,IL-1β and IL-6 were significantly decreased by FC (P＜0.05).Conclusion Astrocytes may play an important role in early brain injury after SAH,underlying the mechanism that they released TNF-α,IL-1βand IL-6 and activate microglia by activating NF-κB signal.The inhibition of excess activation of astrocytes may plays a protective role in early brain injury after SAH.

Objective:To study the effect of Xuebijing on inflammatory response and prognosis in patients with septic shock.Methods:A prospective randomized controlled study was conducted. Eighty septic shock patients admitted to department of critical care medicine of the First Affiliated Hospital of Zhengzhou University from January to December in 2019 were enrolled. The enrolled patients were divided into Xuebijing group and control group by randomized number table method, with 40 cases in each group. Both groups were strictly followed the guidelines for the diagnosis and treatment of septic shock to take comprehensive treatment measures against sepsis. On this basis, Xuebijing group received intravenous 100 mL Xuebijing injection twice a day for 7 days. Baseline data of enrolled patients were recorded. The levels of interleukin-6 (IL-6), procalcitonin (PCT), C-reactive protein (CRP) and heparin binding protein (HBP) were measured before treatment and 3, 7 and 10 days after treatment. Mechanical ventilation time, the length of intensive care unit (ICU) stay, total hospitalization time and 28-day mortality were recorded. The differences of every indicator between the two groups were compared. Independent risk factors affecting patient prognosis were analyzed by binary Logistic regression.Results:① There was no significant difference in baseline data such as gender, age, infection site, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure score (SOFA) between the two groups. ② The levels of serum inflammatory factors in both groups showed a decreasing trend after treatment. Compared with the control group, IL-6 and HBP in the Xuebijing group significantly decreased on day 7 [IL-6 (ng/L): 66.20 (16.34, 163.71) vs. 79.81 (23.95, 178.64), HBP (ng/L): 95.59 (45.23, 157.37) vs. 132.98 (73.90, 162.05), both P < 0.05]; on day 10, PCT, CRP, IL-6 and HBP significantly decreased [PCT (μg/L): 1.14 (0.20, 3.39) vs. 1.31 (0.68, 4.21), CRP (mg/L): 66.32 (19.46, 115.81) vs. 89.16 (20.52, 143.76), IL-6 (ng/L): 31.90 (13.23, 138.74) vs. 166.30 (42.75, 288.10), HBP (ng/L): 62.45 (29.17, 96.51) vs. 112.33 (58.70, 143.96), all P < 0.05]. ③ Compared with the control group, mechanical ventilation time and the length of ICU stay were significantly shortened and the total hospitalization expenses were significantly reduced in Xuebijing group [mechanical ventilation time (hours): 57.0 (0, 163.5) vs. 168.0 (24.0, 282.0), the length of ICU stay (days): 8.80±4.15 vs. 17.13±7.05, the total hospitalization expenses (ten thousand yuan): 14.55±7.31 vs. 20.01±9.86, all P < 0.05]. There was no significant difference in 28-day mortality and the total hospitalization time [28-day mortality: 37.5% vs. 35.0%, the total hospitalization time (days): 13.05±8.44 vs. 18.30±9.59, both P > 0.05]. ④ Patients were divided into death and survival groups according to the prognosis, and univariate analysis showed that white blood cell (WBC), neutrophil percentage (NEU%), CRP, lactic acid (Lac), APACHEⅡ score, IL-6, HBP were the factors influencing the prognosis of patients. The above indicators were further analyzed by Logistic regression, which showed that CRP, IL-6, and APACHE Ⅱ score were independent risk factors for prognosis [odds ratio ( OR) was 1.007, 1.828, 1.229, all P < 0.05]. Conclusions:Combined with Xuebijing to treat septic shock can reduce the body's inflammatory response to a certain extent, thereby reducing the time of mechanical ventilation, shortening the stay of ICU and reducing the total cost of hospitalization. But it cannot reduce the 28-day mortality of patients with septic shock.

Objective:To explore effect of walking on rehabilitation in aged patients with hypertension complicated anxiety.Methods:A total of 160 aged patients with hypertension complicated anxiety were enrolled,randomly divided into medica-tion group (n＝80,received routine medication ) and exercise group (n＝80,received walking exercise based on routine medication),both groups were treated for four months.Carotid intima-media thickness (IMT) and ZUNG self-rating anxi-ety scale (SAS) score before and after treatment,incidence rate of cardiovascular events after one-year follow-up were compared between two groups.Results:Compared with before treatment,there were significant reductions in IMT and SAS score in two groups after treatment,P＝0.001 all;compared with medication group after treatment,there were significant reductions in IMT [ (1.05 ± 0.26) mm vs.(0.90 ± 0.46) mm] and SAS [ (38.07 ± 0.25) scores vs.(23.02 ± 0.46) scores] in exercise group,P＜0.01 both.During one-year follow-up,total incidence rate of cardiovascular events in exer-cise group was significantly lower than that of medication group (30.67% vs.69.74%,P＝0.001).Conclusion:Walking helps to improve anxiety,reduce incidence of adverse cardiovascular events in aged patients with hypertension complicated anxiety.It’ s safe and reliable,which is worth extending.